Murine dorsal skins were embedded in OCT compound (Tissue-Tek; American Master Tech Scientific, Lodi, CA), frozen, and sectioned. Tissue cross-sections were blocked with 10% normal goat serum and immunostained with antibodies for CD34 (1:100, BD Pharmingen, Franklin Lakes, NY), CDK4 (1:50, Santa Cruz Biotech, Dallas, TX), and anti-GFP (1:50, abcam1218, Abcam, Burlingame, CA) followed by incubation with Alexafluor secondary antibodies (FITC or Texas re-conjugated anti-Rat or anti-goat; Molecular Probes, Eugene, OR). Frozen cross-sections were counterstained with 4’6’-diamidino-2-phenylindole (DAPI) and visualized under a fluorescence microscope using a 465 to 495 nm filter. BrdU incorporation was detected by immunohistochemical staining of paraffin-embedded skin sections with mouse anti-BrdU (Ab-2) monoclonal antibody (Calbiochem, EMB Biosciences, San Diego, CA), biotin-conjugated anti-mouse antibody (Vector Laboratories, Inc.), and an avidin-biotin-peroxidase kit (Vectastain Elite, Vector Laboratories) with diaminobenzidine as the chromogen.
Abcam1218
Manufactured by Abcam
Abcam1218 is a laboratory equipment product. It serves a core function, but a detailed description while maintaining an unbiased and factual approach is not available.
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2 protocols using abcam1218
1
Murine Skin Immunohistochemistry Protocol
2
Immunohistochemical Analysis of Murine Skin
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Murine dorsal skins were embedded in OCT compound (Tissue‐Tek; American Master Tech Scientific), frozen, and sectioned. Tissue cross‐sections were blocked with 10% normal goat serum and immunostained with antibodies for CD34 (1:100, BD Pharmingen), CDK4 (1:50, Santa Cruz Biotech), and anti‐GFP (1:50, abcam1218, Abcam) followed by incubation with Alexafluor secondary antibodies (FITC or Texas re‐conjugated anti‐Rat or anti‐goat; Molecular Probes). Frozen cross‐sections were counterstained with 4ʹ6ʹ‐diamidino‐2‐phenylindole (DAPI) and visualized under a fluorescence microscope using a 465–495 nm filter. BrdU incorporation was detected by immunohistochemical staining of paraffin‐embedded skin sections with mouse anti‐BrdU (Ab‐2) monoclonal antibody (Calbiochem, EMB Biosciences), biotin‐conjugated anti‐mouse antibody (Vector Laboratories, Inc.), and an avidin‐biotin‐peroxidase kit (Vectastain Elite, Vector Laboratories) with diaminobenzidine as the chromogen.
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